A missense polymorphism in porcine interferon-γ cDNA affects antiviral activity of the protein variant
| Autor: | Shiow-Her Chiou, Lang-Ming Chi, Kuan-Chih Chow, Chienjin Huang, San-Yuan Huang, Yi-Hsin Fan |
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| Rok vydání: | 2007 |
| Předmět: |
DNA
Complementary Swine Molecular Sequence Data Immunology Mutation Missense Single-nucleotide polymorphism Viral Plaque Assay Biology medicine.disease_cause Antiviral Agents Article law.invention Interferon-gamma law Complementary DNA medicine Animals Humans Coding region Interferon gamma Amino Acid Sequence Polymorphism Antiviral activity Molecular Biology Escherichia coli Peptide sequence Cells Cultured Porcine IFN-γ Genetics Polymorphism Genetic Base Sequence Nucleic acid sequence Genetic Variation Herpesvirus 1 Suid Molecular biology Functional SNP Recombinant DNA Virus Inactivation medicine.drug |
| Zdroj: | Molecular Immunology |
| ISSN: | 0161-5890 |
| DOI: | 10.1016/j.molimm.2007.02.029 |
| Popis: | We determined the interferon-gamma (IFN-gamma) cDNA sequence from three porcine breeds, Duroc, Landrance/Duroc hybrid, and Landrance breeds. Five single nucleotide polymorphisms (SNPs) of porcine IFN-gamma (PoIFN-gamma) were identified, respectively, at positions 269 (A/G), 376 (C/T), 426 (T/C), and 465 (T/C) of the coding sequence in Landrance/Duroc hybrid, and at position 251 (A/G) in Landrance breed. Among them, A269G and A251G polymorphisms resulted in Q67R and K61R replacements in the mature protein. PoIFN-gamma cDNAs of Duroc breed (PoIFN-gamma-W) and Landrance/Duroc hybrid (PoIFN-gamma-M), which, respectively, encoded Q67 and R67, were introduced into a prokaryotic expression vector pET32 to express recombinant PoIFN-gamma-W (rPoIFN-gamma-W) and rPoIFN-gamma-M protein variants in Escherichia coli. The identity of both protein variants was further confirmed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). We then compared bioactivities of these two recombinant proteins. Although both recombinant protein variants exhibited comparable activities in antiproliferation of PK-15 cells and in nitric oxide (NO) induction of porcine peripheral monocytes, antiviral activity of rPoIFN-gamma-W protein was significantly higher (P |
| Databáze: | OpenAIRE |
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